[Histonet] Fwd: spinal cord paraffin embedding

Elisa Ballarini e.ballarini1 <@t> campus.unimib.it
Tue Mar 16 10:33:54 CDT 2010


   Hi.
   First of all I would like to thank all of you for your advices.
   Because,  my  previous  e-mail  wasn't  complete  and to avoid further
   suggestions  regarding  protocols  I can not follow, here I write some
   more information.
   First  of  all,  we  can't perfuse our rats. Also, when we harvest rat
   spinal cord (cut in half, thus around 3mm diameter) we fix it only for
   4-5  hours  in  4%  paraformaldehyde because we need to perform an IHC
   essay so we would like to avoid epitope masking.
   We  don't  think  fixation is the problem since frozen sections of the
   same fixed spinal cords do not show any problem in the white matter.
   In  conclusion,  we need to obtain good results using paraffin both in
   term   Our paraffin embedding machine program is as follows:
   Ethanol 95%   1h
   Ethanol 95%   45'
   Ethanol 95%   45'
   Ethanol 100%  45'
   Ethanol 100%  1h
   Ethanol 100%  1h
   Ethanol 100%  1h
   Xylol 45'
   Xylol 1h
   Xylol 1h
   Paraffin 1h
   Paraffin 1h
   Paraffin 1h
   Can  you  tell  me  if  are there any specific protocols to embedd rat
   spinal    Thank you.
   Regards
   --
   -- 
   Dott.ssa Elisa Ballarini,PhD student
   Dipartimento di Neuroscie   Università degli Studi Milano-Bicocca
   Via Cadore 48-20052,Monza,MB
   e.ballarini1 <@t> campus.uni   Tel. 02-64488119
   Fax. 02-64488253

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